DISINFECTANT

CONTENT

Terminology
Introduction of disinfectants
Classification of disinfectants
Mode of action of disinfectants
Factors affecting disinfectants
Evaluation of disinfectants
 TERMINOLOGY

• Sepsis: Bacterial contamination.

• Asepsis: Absence of significant contamination.
• Antisepsis: Chemical destruction of vegetative
pathogens on living tissue.
• Sanitization: Lowering microbial counts on eating
and drinking utensils to safe level.
 TERMINOLOGY

• Bactericidal: Chemical agents capable of killing bacteria.

• Virucidal: Chemical agents capable of killing bacteria.
• Fungicidal: Chemical agents capable of killing fungi.
• Sporicidal: Chemical agents capable of killing spores.
• Bacteriostatic: Chemical agents that inhibit the growth
of bacteria but do not necessarily kill them.
 INTRODUCTION OF DISINFECTANTS

• Disinfection is a process of destruction or removal of microorganisms and

reducing them to the level not harm to health.
• Disinfectant generally kill the sensitive vegetative cells but not heat resistance
endospores.
• If the object is inanimate (lifeless), such as working area, dishes, benches, etc. the
chemical agent is known as disinfectants.
• However if the object is animate (living) such as human body tissue, the chemical is
known as antiseptic.
• Disinfectants are usually bactericidal but occasionally they may be bacteriostatic.
 AN IDEAL DISINFECTANT SHOULD HAVE
FOLLOWING PROPERTIES.
• Broad spectrum
• Non toxic
• Fast acting
• Odourless
• Surface compatibility
• Economical
• Easy to use
• Soluble and miscibility
• Not affected by the physical factors
• Stable on storage
 CLASSIFICATION OF DISINFECTANTS

The chemical agents are classified as follows:

1. Acid and alkalies
2. Halogens
3. Heavy metals
4. Phenol and its derivatives
5. Alcohols
6. Quaternary ammonium compounds
7. Dyes
8. Detergents and Soaps
9. Aldehydes
1. Acids and alkalies
Generally, strong acids and alkalis kill the bacteria but weak organic acids inhibit
their growth.
2. Halogens
Chlorine, fluorine, bromine and iodine in the free state as well as their
compounds strongly act as germicidal.
3.Heavy metals
Mercury, silver and copper (widely used heavy metals)
High concentration of heavy metals coagulate cytoplasmic proteins, resulting the
damage or death of cell.
4.Phenol and its derivatives
Phenol is the chief product obtained by distillation of the coal tar.
Phenol 1% has bactericidal action.
Many derivatives of phenol are more effective and less costly.
5. Alcohols
Alcohols have fairly rapid bactericidal action against
vegetative bacteria when diluted to the concentration of
60% to 70% v/v with water.
The higher alcohols (propyl, butyl, amyl etc) are more
germicidal than ethyl alcohol.
Alcohols are used as preservatives in some vaccines.
6. Aldehyde
Formaldehyde (HCHO) is the main aldehyde used for
disinfection.
It is also useful for sterilization of certain instrument.
7. Quaternary ammonium compound
Quaternary ammonium compound are widely used for the
control of microorganism on floors, walls, nursing homes, and
other public places.
They are also used as skin antiseptics and as sanitizing
agents in dairy, egg and fishing industries.
8.Dyes
A number of dyes have been used to inhibit the bacterial
growth.
9.Detergents and soaps
They are are widely used as surface active agents, wetting
agents and emulsifiers.
 MODE OF ACTION OF DISINFECTANTS

Alternation of membrane permeability.

Damage to protein.
Rupture of cell membrane.
Damage to nucleic acids.
Interfere with metabolic pathway.
 FACTORS AFFECTING DISINFECTANTS
i. Concentration of disinfectants
ii. Temperature
iii. Time of contact
iv. ph of environment
v. Surface tension
vi. Formulation of disinfectants
vii. Chemical structure of disinfectants
viii. Types and number of microorganism present
ix. Interfering substances in the environment
 EVALUATION OF ANTI-MICROBIAL AGENTS
AND DISINFECTANTS

Techniques and methods used for evaluation of

disinfectants
Tube dilution and agar plate method
Cup plate method or Cylinder plate method
Ditch plate method
Gradient plate technique
Phenol coefficient method (Rideal-Walker test)
 1. TUBE DILUTION AND AGAR PLATE
METHOD

• The chemical agent is incorporated into nutrient broth or

agar medium and inoculated with test micro-organism.
• These tubes are incubated at 30C to 35C for 2 to 3 days
and then the results in the form or turbidity or colonies are
observed.
• The results are recorded and the activity of given
disinfectant is compared.
2.CUP PLATE OR CYLINDER PLATE METHOD

• The nutrient agar is melted, cooled suitably, poured into petri dish.
• Spread 0.2 ml of known concentration of inoculum on the surface of
the solidified agar (Spread Plate Technique).
• Cups or cavities are made by using a sterile borer.
• Now 0.2ml of drug is poured into the cups of agar plate and then
incubated at 37C for 24 hours.
• If the drug has any anti-bacterial effect it will show the zone of
inhibition.
 3.DITCH-PLATE METHOD

 The nutrient agar is melted, cooled suitably , poured into petri dish.
 The solidified media is cut with a sterile blade to make a ditch.
 The drug is poured vary carefully into the ditch.
 Various microorganism are streaked on the slide of the ditch.
 This method is used to find out the potency of drug against various
microorganisms by the means of inhibition of growth on streaked
area.
 4. GRADIENT PLATE TECHNIQUE
• This technique is used to isolate the resistance mutants.
• The petri dish is kept in slanting position and a sufficient amount of melted
nutrient agar is poured and solidified in slanting position.
• Another layer of agar is poured over it, which contains antibiotic solution and
solidified it.
• After solidification, 0.2ml of bacterial culture was spreaded over the solid
surface and incubated it at 37C for 24 to 48 hours.
• The microorganism will grow, where the concentration of the drug is below
the critical level.
• The antibiotic get diluted on the lower layer and the gradient of concentration
will be produced.
• Thus the resistant mutant can b isolated.
5. PHENOL COEFFICIENT METHOD(RIDEAL-
WALKER TEST)

• Phenol coefficient test is suitable for testing

disinfectants miscible with water and which
exert their anti microbial action in manner similar
to that of phenol.
• Test organism: Salmonella typhi
• Standard disinfectant: Phenol
How do you measure the efficacy of a disinfectant?

The disc diffusion method is used to test the effectiveness of a chemical

disinfectant against a particular microbe. The use-dilution test determines
the effectiveness of a disinfectant on a surface. In use tests can
determine whether the disinfectant solution are being used correctly in
clinical settings.

How effective disinfectants are?

Disinfectants does not necessarily kill all microorganisms, especially

resistant bacterial spores, it is less effective than sterilization, which is an
extreme physical or chemical process that kill all type of life. Disinfectants
work by destroying the cell wall of microbes or interfering with their
metabolism.